Puerarin, an isoflavone extracted from Radix puerariae, is a 5-HT2C receptor antagonist.

Puerarin inhibits the expression of LPS-induced iNOS, COX-2 and CRP proteins and also suppresses their mRNAs from RT-PCR experiments in RAW264.7 cells. The inhibition of iNOS, COX-2 and CRP expression is due to a dose-dependent inhibition of phosphorylation and degradation of I-κB, which resulted in the reduction of p65NF-κB nuclear translocation. The effect of puerarin-mediated inhibition of LPS-induced iNOS, COX-2 and CRP expression is attributed to suppressed NF-κB activation at the transcriptional level^[1]. Puerarin is a novel open-channel blocker of IK1, which may underlie the antiarrhythmic action of puerarin. Puerarin competes with barium, an open-channel blocker of IK1, to inhibit IK1 currents^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Both genistein and puerarin effectively alleviate hepatic damage induced by chronic alcohol administration through potential antioxidant, anti-inflammatory, or anti apoptotic mechanisms. However, genistein is more effective than puerarin in decreasing levels of malondialdehyde (1.05±0.0947 vs. 1.28±0.213 nmol/mg pro, p< 0.05), tumor necrosis factor α (3.12±0.498 vs. 3.82±0.277 pg/mg pro, p < 0.05), interleukin-6 (1.46±0.223 vs. 1.88±0.309 pg/mg pro, p < 0.05), whereas puerarin is more effective than genistein in ameliorating serum activities or levels of alanine transaminase (35.8±3.95 vs. 42.6±6.56 U/L, p < 0.05) and low-density lipoprotein cholesterol (1.12±0.160 vs. 1.55±0.150 mmol/L, p < 0.05) ^[3]. Early-stage renal damages can be significantly improved by puerarin, possibly via its suppression of ICAM-1 and TNF-α expression in diabetic rat kidneys^[4].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

416.38

C₂₁H₂₀O₉

3681-99-0

葛根素；葛根黄酮

Room temperature in continental US; may vary elsewhere.

DMSO : 50 mg/mL (120.08 mM; Need ultrasonic)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂：;20% SBE-β-CD in saline

  Solubility: 20 mg/mL (48.03 mM); Clear solution; Need ultrasonic
• 2.

  请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 3 mg/mL (7.20 mM); Clear solution

  此方案可获得 ≥ 3 mg/mL (7.20 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 30.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 3.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 3 mg/mL (7.20 mM); Clear solution

  此方案可获得 ≥ 3 mg/mL (7.20 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 30.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 4.

  请依序添加每种溶剂：;10% DMSO ;; 90% corn oil

  Solubility: 3 mg/mL (7.20 mM); Suspended solution; Need ultrasonic

  此方案可获得 3 mg/mL (7.20 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 30.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Hu W, et al. Puerarin inhibits iNOS, COX-2 and CRP expression via suppression of NF-κB activation in LPS-induced RAW264.7 macrophage cells. Pharmacol Rep. 2011;63(3):781-9.

  [2]. Zhang H, et al. Puerarin: a novel antagonist to inward rectifier potassium channel (IK1). Mol Cell Biochem. 2011 Jun;352(1-2):117-23.

  [3]. Zhao L,et al. Protective Effects of Genistein and Puerarin against Chronic Alcohol-Induced Liver Injury in Mice via Antioxidant, Anti-inflammatory, and Anti-apoptotic Mechanisms. J Agric Food Chem. 2016 Sep 28;64(38):7291-7.

  [4]. Pan X, et al. Effect of Puerarin on Expression of ICAM-1 and TNF-α in Kidneys of Diabetic Rats. Med Sci Monit. 2015 Jul 23;21:2134-40.

RAW264.7 cells are maintained at subconfluence in 95% air and 5% CO₂ humidified atmosphere maintained at 37°C. The medium used for routine subculture is Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum, penicillin (100 units/mL) and streptomycin (100 μg/ mL). An MTT assay is used to measure the viability of the cells after treatment with puerarin. After the supernatants are removed for nitrite determination, cells are incubated at 37°C with MTT (0.05 mg/mL) for 4 h, and the optical density is measured at 540 nm. The concentrations of puerarin are10, 20, 40 and 100 μM^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Rats: A cohort of healthy male SD rats (7 weeks old) are randomLy divided into a control group, a model group, and a puerarin treatment group with high (H), moderate (M), and low (L) dosage. Puerarin is re-suspended in 0.9% saline and is given by intra-gastric intubation at various concentrations (0.25 mg/(kg×d) for L group, 0.5 mg/(kg×d) for M group, and 1.0 mg/(kg×d) for H group) each day for 8 consecutive days. An equal volume of saline is administered to control and model rats during the same time period^[4].

Mice: Forty male ICR mice (weight: 20-22 g) are acclimatized with a daily 12 h light:12 h dark cycle at 22±2 °C room temperature and 55%±5% relative humidity. After 1 week of adaption, the mice are randomLy divided into four groups with ten mice per group. Genistein and puerarin are applied to the mice in sodium carboxymethyl cellulose solution with an equimolar concentration of 0.1 M (gastric volume: 3 mL kg^-1 body weight)^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Hu W, et al. Puerarin inhibits iNOS, COX-2 and CRP expression via suppression of NF-κB activation in LPS-induced RAW264.7 macrophage cells. Pharmacol Rep. 2011;63(3):781-9.

  [2]. Zhang H, et al. Puerarin: a novel antagonist to inward rectifier potassium channel (IK1). Mol Cell Biochem. 2011 Jun;352(1-2):117-23.

  [3]. Zhao L,et al. Protective Effects of Genistein and Puerarin against Chronic Alcohol-Induced Liver Injury in Mice via Antioxidant, Anti-inflammatory, and Anti-apoptotic Mechanisms. J Agric Food Chem. 2016 Sep 28;64(38):7291-7.

  [4]. Pan X, et al. Effect of Puerarin on Expression of ICAM-1 and TNF-α in Kidneys of Diabetic Rats. Med Sci Monit. 2015 Jul 23;21:2134-40.